Imaging B. anthracis heme catabolism in mice using the IFP1.4 gene reporter

Banghe Zhu; Holly Robinson; Nathaniel Wilganowski; Christopher Loren Nobles; Eva Sevick-Muraca; Anthony Maresso

doi:10.1117/12.909061

2 February 2012 Imaging B. anthracis heme catabolism in mice using the IFP1.4 gene reporter

Banghe Zhu, Holly Robinson, Nathaniel Wilganowski, Christopher Loren Nobles, Eva Sevick-Muraca, Anthony Maresso

Proceedings Volume 8233, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications IV; 82330K (2012) https://doi.org/10.1117/12.909061
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

B. anthracis is a gram-positive, spore-forming bacterium which likes all pathogenic bacteria, survive by sequestering heme from its host. To image B. anthracis heme catabolism in vivo, we stably transfect new red excitable fluorescent protein, IFP1.4, that requires the heme catabolism product biliverdin (BV). IFP1.4 reporter has favorable excitation and emission characteristics, which has an absorption peak at 685 nm and an emission peak at 708 nm. Therefore, IFP1.4 reporter can be imaged deeply into the tissue with less contamination from tissue autofluorescence. However, the excitation light "leakage" through optical filters can limit detection and sensitivity of IFP1.4 reporter due to the small Stoke's shift of IFP1.4 fluorescence. To minimize the excitation light leakage, an intensified CCD (ICCD) based infrared fluorescence imaging device was optimized using two band pass filters separated by a focus lens to increase the optical density at the excitation wavelength. In this study, a mouse model (DBA/J2) was first injected with B. anthracis bacteria expressing IFP1.4, 150 μl s.c., on the ventral side of the left thigh. Then mouse was given 250 μl of a 1mM BV solution via I.V. injection. Imaging was conducted as a function of time after infection under light euthanasia, excised tissues were imaged and IFP1.4 fluorescence correlated with standard culture measurements of colony forming units (CFU). The work demonstrates the use of IFP1.4 as a reporter of bacterial utilization of host heme and may provide an important tool for understanding the pathogenesis of bacterial infection and developing new anti-bacterial therapeutics.

Citation Download Citation

Banghe Zhu, Holly Robinson, Nathaniel Wilganowski, Christopher Loren Nobles, Eva Sevick-Muraca, and Anthony Maresso "Imaging B. anthracis heme catabolism in mice using the IFP1.4 gene reporter", Proc. SPIE 8233, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications IV, 82330K (2 February 2012); https://doi.org/10.1117/12.909061

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KEYWORDS

Luminescence

In vivo imaging

Tissues

Pathogens

Heart

Optical filters

Fluorescent proteins

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